Migraine is a complex, hereditary, sexually-dimorphic neurological disorder that is a major health problem, particularly for reproductive-aged women. The incidence of the disease increases dramatically in females after puberty, however the cause of this increase still remains elusive. One potential possibility is a genetic predisposition to certain phenotypic changes in the brain that emerges during puberty. Indeed, developmental changes in the brain during the pubertal phase may create individual sex-specific variation in disease evolution trajectory. In migraine disease, a number of studies have pointed to the presence of sex-specific abnormalities in the adult female brain. It is likely that some of these changes may have appeared during puberty, since changes in the brain during puberty are region specific, with some regions changing more dramatically at each stage of growth compared to the others. Our preliminary studies particularly point to two potential sites of susceptibility in the brain that may play a major role in migraine pathophysiology, insula and brainstem. These regions are also implicated by multiple studies reporting structural, functional and functional connectivity abnormalities associated with migraine. Our preliminary studies have suggested: i) female-specific plastic changes in the insula in the adult migraine brain relative to healthy female subjects that differ from healthy and migraineur male subjects; ii) abnormal pattern of no thinning of the gray matter in the insular cortex of adult female migraineurs; iii) sex-related differences in the rate of the development of the brainstem during earlier stages of pubertal development and iv) association of earlier age at menarche with risk of migraine (but not other types of headaches) in adult women. Since ovarian steroid hormone secretion is initiated and governed by genetic mechanisms that control the `timing' of the initiation of puberty, we hypothesize that there may be a link between genetic mechanisms of puberty and risk of migraine in adulthood in women. A recent meta-analysis of genome-wide association studies (GWAS) has identified over 100 genomic loci that influence age at menarche, a marker of pubertal timing in females. In Aim 1, we propose to determine the association of menarche genetic loci with migraine risk in a large female cohort (~48,000 patients, ~150,000 controls). The identification of significant associations will provide compelling evidence for molecular links between pubertal development and migraine risk in females. In Aim 2, we propose to examine the relationship between migraine and structural brain changes in female patients by analyzing a large set of legacy clinical MRI data (1500 patients, 900 controls). In a subset of these patients (315 patients, 420 controls), we also propose to explore the association between menarche genetic markers and brain structural changes that are related to migraine. Identifying molecular links between migraine disease and pubertal development may allow for a better understanding of the mechanisms underlying disease onset and progression that will provide critical information for the discovery of novel drug targets for modifying the course of migraine in susceptible individuals.